1.Prescription parsing of miao medicine Polygonum capitatum and kelintong capsule.
Nan HU ; Shi-Huan TANG ; Long CHENG
China Journal of Chinese Materia Medica 2014;39(7):1318-1320
By literature survey searching references and parsing prescriptions, the auther has analyze the clinical advantage of Miao medicine in the treatment of symptom heat stranguria. Guizhou Miao medicine Polygonum capitatum has many advantages such as resources and clinical. After companying with Phellodendri Cortex, the compound prescription plays the pharmacological activity of antipyretic and diuretic, especially for the symptom heat stranguria, damp and hot junction based in the bladder. Miao medicine Kelintong capsule showed clinical advantage in the treatment of symptom heat stranguria, having a clinical advantage in improving the overall effectiveness and improve the overall aspects of the patient's symptoms.
Animals
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Capsules
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chemistry
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therapeutic use
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Drug Prescriptions
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Drugs, Chinese Herbal
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chemistry
;
therapeutic use
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Humans
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Medicine, Chinese Traditional
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Polygonum
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chemistry
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Urination Disorders
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drug therapy
2.The development of the measuring equipment for rotating speed and temperature of the centrifuge
Huan CHENG ; Junfu SHI ; Jianhua JIANG ; Ying HUANG
Chinese Medical Equipment Journal 1989;0(01):-
To measure the rotating speed and temperature of the centrifuge, an equipment and method are developed in this paper. They prove to meet the requirements of measuring. The equipment and method can be used to measure the rotating speed or temperature of the operating centrifuge easily and accurately. That is also the main excellence of them.
3.mRNA expression change of myelin gene in hippocampus following cerebral ischemia
Ying-Zhu CHEN ; Huan BAO ; Ye TIAN ; Shi-Yao BAO ; Jun XU ; Cheng-Lin YUAN ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To study the mRNA expression of myelin basic protein (MBP) and myelin oligodendregha glyeoprotein (MOG) in hippocampus of rats following global brain ischemia.Method The four- vessel occlusion animal model in the Sprague-Dawley rats was used in this study.The mRNA expression levels of MBP and MOG in the hippocampus of rats were analyzed by reverse transcription polymerase chain reaction (RT- PCR) at day 2,4,7,14 and 28 days after global brain ischemia.There were eight rats at each time-point and sham operated group.Results The mRNA expression of both MBP and MOG in hippocampus of rats decreased at 2 days after global brain ischemia.The gene expression of myelin gene decreased significantly at 7 days and it reached to the lowest level at 28 days.Compared with sham operated group,the gene expression of MBP and MOG in hippocampus of rats decreased significantly at 7,14 and 28 days after global brain ischemia (P
5.Mechanism of BCL2L2-PABPN1 expression induced by sodium arsenite and its metabolites in 16HBE cells
SHI Ya YIN Jin yao WU Jiang JIANG Cheng lan ZHAO Rui huan ZHOU Qian HE Yue feng
China Occupational Medicine 2022;49(05):522-
Objective - - (BCL2L2)- ( )
To investigate the differential expression of the fusion gene BCL 2 like protein 2 poly A
(PABPN1) ( )
binding protein nuclear 1 induced by sodium arsenite SA and its methylated metabolites in 16HBE cells and the
Methods ) ,
related mechanism. i The 16HBE cells exposed to SA at concentrations of 1.5 3.0 and 4.5 µmol/L were set as
-, - -
low medium and high dose arsenic exposure groups. The 16HBE cells exposed to 4.5 µmol/L monomethylarsonic acid
( ), ( ) ,
MMA dimethylarsonic acid DMA and SA were set as MMA group DMA group and SA group. The 16HBE cells without
, BCL2L2-PABPN1
toxic stimulation were set as control group. After the cells were cultured for 48 hours the expression of was
- ( - ) ) ( )
detected by quantitative real time polymerase chain reaction qRT PCR . ii Two small interfering RNA siRNA silencing
基金项目:国家自然科学基金( ); 年云南省科技厅昆明医科大学应用基础研究联合专项面上项目
82160607 2021
( )
202101AY070001-054
作者简介:施雅( —),女,在读大学本科生,主要从事劳动卫生与环境卫生学研究;尹锦瑶( —),女,在读劳动卫生与环境卫
2001 1995
生学硕士研究生,主要从事劳动卫生与环境卫生学研究;施雅和尹锦瑶为共同第一作者
通讯作者:何越峰教授,博士研究生导师,- :
E mail heyuefeng@kmmu.edu.cn中国职业医学 年 月第 卷第 期 , , , · ·
2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 523
BCL2L2-PABPN1, -
fragments were designed and transfected into 16HBE cells to knockdown which were set as siRNA 1 group
- - BCL2L2-PABPN1
and siRNA 2 group. Non transfected control group without knockdown of transfection was set up. After
, BCL2L2-PABPN1 -
culturing for 48 hours the expression level of in the three groups of cells was detected by qRT PCR. The cell
-
survival rate and early apoptosis rate were detected by MTS method and JC 1 mitochondrial membrane potential detection
, ( ) ,
method respectively. The apoptosis was detected by Hoechest33342/propidium iodide PI double staining and the expression
- Results )
level of P53 signaling pathway related proteins was detected by Western blotting. i The relative expression of
BCL2L2-PABPN1 (P ) BCL2L2-
in 16HBE cells increased with the increasing SA doses <0.01 . The relative expression of
PABPN1 - , - -
in high dose arsenic exposure was higher than that in control group low dose and medium dose arsenic exposure
( P ) BCL2L2-PABPN1 ,
groups all <0.05 . The relative expression of in SA group was higher than those in control group MMA
( P ) BCL2L2-PABPN1
group and DMA group all <0.05 . The relative expression of showed no significant difference between
, ( P ) ) BCL2L2-PABPN1
control group MMA group and DMA group all >0.05 . ii The relative expression levels of and cell
- - - ( P )
survival rate in siRNA 1 group and siRNA 2 group were lower than those in non transfected control group all <0.05 .
, (P )
However there was no significant difference in the early apoptosis rate among the three groups >0.05 . The results of
-
Hoechest33342/PI double staining showed that the number of nuclear shrinkage and early apoptotic cells in siRNA 1 group and
- - , -
siRNA 2 group was higher than that in non transfected control group. The relative protein expression levels of P53 phospho
, - - , - - ( P )
p53 BCL 2 associated death promoter P21 and cytochrome C in siRNA 1 group and siRNA 2 group were higher all <0.05 ,
- - P
and the relative protein expression levels of P53 up regulated modulator of apoptosis were lower (all <0.05), when compared
- Conclusion
with the non transfected control group. SA may block the apoptosis of 16HBE cells by inducing the expression of
BCL2L2-PABPN1
fusion gene . The mechanism may be related to the activation of P53 signaling pathway. The SA methylated
BCL2L2-PABPN1 BCL2L2-PABPN1 -
metabolites MMD and DMA had no effect on the expression of . may affect anti apoptosis
BCL2L2 PABPN1
through affecting the synergistic effect of and genes.
6.Construction and identification of recombinant human platelet-derived growth factor-B adenoviral vector and transfection into periodontal ligament stem cells
Shu-Huan SHANG ; Yu-Feng ZHANG ; Bin SHI ; Xiang-Rong CHENG
Chinese Journal of Stomatology 2008;43(10):584-588
Objective To construct a recombinant human platelet-derived growth factor-B (PDGF-B) adenoviral vector and to transfect it into human periodontal ligament stem cells (PDLSC).Methods The recombinant plasmid pAd-PDGF-B was constructed by homologous recombination and confirmed by restriction endonucleases digestion.Recombinant adenovirus was packaged in HEK293 cells.PDLSC were transfected with recombinant adenovirus and PDGF-B expression was confirmed.Expression of collagen type Ⅰ gene was determined by quantitative analysis of the products of RT-PCR.The cell proliferation was determined with MTr eolorimetric assay.Results The recombinant plasmid pAd-PDGF-B was confirmed by restriction endonucleases digestion.EGFP expression was observed on the third day after transfecting,and the expression of PDGF-B was detected.Immunohistochemical methods revealed that PDGF-B was expressed in PDLSC.Levels of expression of collagen type Ⅰ gene were increased significantly by transfer of the exogenous PDGF-B gene to PDLSC.At the same time,findings indicated that Ad-PDGF-B stimulated PDLSC proliferation.MTT assay indicated the absorbance of PDLSC by stimulating with Ad-PDGF-B was (0.68±0.02),P<0.01.Conclusions Using the AdEasy system,the human PDGF-B recombinant adenovirns can be rapidly obtained.These results indicate that recombinant adenoviruses encoding PDGF-B transgenes could modulate proliferative activity of PDLSC,enhance the high expression of collagen type Ⅰ and lay the foundation for periodontal tissue regeneration and dental implant gene therapy.
7.Derivation and characterization of Chinese human embryonic stem cell line with high potential to differentiate into pancreatic and hepatic cells.
Cheng SHI ; Huan SHEN ; Wei JIANG ; Zhi-Hua SONG ; Cheng-Yan WANG ; Li-Hui WEI
Chinese Medical Journal 2011;124(7):1037-1043
BACKGROUNDHuman embryonic stem cells have prospective uses in regenerative medicine and drug screening. Every human embryonic stem cell line has its own genetic background, which determines its specific ability for differentiation as well as susceptibility to drugs. It is necessary to compile many human embryonic stem cell lines with various backgrounds for future clinical use, especially in China due to its large population. This study contributes to isolating new Chinese human embryonic stem cell lines with clarified directly differentiation ability.
METHODSDonated embryos that exceeded clinical use in our in vitro fertilization-embryo transfer (IVF-ET) center were collected to establish human embryonic stem cells lines with informed consent. The classic growth factors of basic fibroblast growth factor (bFGF) and recombinant human leukaemia inhibitory factor (hLIF) for culturing embryonic stem cells were used to capture the stem cells from the plated embryos. Mechanical and enzymetic methods were used to propagate the newly established human embryonic stem cells line. The new cell line was checked for pluripotent characteristics with detecting the expression of stemness genes and observing spontaneous differentiation both in vitro and in vivo. Finally similar step-wise protocols from definitive endoderm to target specific cells were used to check the cell line's ability to directly differentiate into pancreatic and hepatic cells.
RESULTSWe generated a new Chinese human embryonic stem cells line, CH1. This cell line showed the same characteristics as other reported Chinese human embryonic stem cells lines: normal morphology, karyotype and pluripotency in vitro and in vivo. The CH1 cells could be directly differentiated towards pancreatic and hepatic cells with equal efficiency compared to the H1 cell line.
CONCLUSIONSThis newly established Chinese cell line, CH1, which is pluripotent and has high potential to differentiate into pancreatic and hepatic cells, will provide a useful tool for embryo development research, along with clinical treatments for diabetes and some hepatic diseases.
Cell Differentiation ; physiology ; Cell Line ; Embryonic Stem Cells ; cytology ; Hepatocytes ; cytology ; metabolism ; Humans ; Karyotyping ; Pancrelipase ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
8.Case-control study on bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle joint.
Yue-Feng QI ; Fa-Lin CHEN ; Shu-Ren BAO ; Cheng-Huan LI ; Xing-Wei ZHAO ; Shi-Ming LIU ; Wen-Xue CHEN ; Ye LI ; Peng WANG
China Journal of Orthopaedics and Traumatology 2012;25(8):634-638
OBJECTIVETo explore therapeutic effects of bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle,and analyze manipulative reduction mechanism.
METHODSFrom 2005 to 2008, 95 patients with over degree II supination-eversion fractures of ankle were treated respectively by manipulation and operation. There were 43 cases [11 males and 32 females with an average age of (44.95 +/- 12.65) years] in manipulation group, and 2 cases were degree II, 11 cases were degree III, and 30 cases were degree IV. There were 52 cases [21 males and 31 females with an average age of (39.96 +/- 13.28) years] in operative group,and 6 cases were degree II, 18 cases were degree III, and 28 cases were degree IV. Bone setting manipulation and hard splint external fixation were applied to manipulative group. Operative reduction internal fixation was performed in operative group. X-ray was used to evaluate reduction of fracture before and after treatment, 2 months after treatment. Ankle joint function was evaluated according to Olerud-Molander scoring system after 6 months treatment.
RESULTSAll patients were followed up with good reduction. Three cases occurred wound complication in operative group, but not in manipulative group. In manipulation group, 19 cases got excellent results, 20 cases good and 4 cases fair; while in operative group, 30 cases got excellent results, 20 cases good and 2 cases poor. There were no significant differences in fracture reduction and ankle joint function recovery between two groups (P > 0.05). Efficacy of operative treatment was better than that of manipulative treatment at degree IV fracture (P < 0.05).
CONCLUSIONBone setting manipulation is a good method for treating supination-eversion ankle joint fractures, which has advantages of simple and safe operation, reliable efficacy. For ankle join fracture at degree IV, manipulative reduction should be adopted earlier, and operative treatment also necessary
Adult ; Ankle Injuries ; physiopathology ; therapy ; Ankle Joint ; physiology ; Case-Control Studies ; Female ; Fractures, Bone ; physiopathology ; therapy ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Supination
9.Cytotoxic effects of mono-(2-ethylhexyl) phthalate on human embryonic stem cells.
Cheng SHI ; Xi CHEN ; Xiao-hui CAI ; Wei-dong YU ; Rong LIANG ; Qun LU ; Huan SHEN
Chinese Medical Journal 2013;126(9):1714-1719
BACKGROUNDMono-(2-ethylhexyl) phthalate (MEHP), the metabolite of di-(2-ethylhexyl) phthalate (DEHP), was suspected to be toxic to human embryos. This study contributes to investigating its toxic effects by an embryonic stem cell test (EST) based on two human embryonic stem cell (hESCs) lines.
METHODSCH1 established in our own lab and H1, a federally registered cell line were two human embryonic stem cell lines used in this test. Four endpoint measurements were performed consisting of cell viability, proliferation ability, apoptosis as well as changes of gene expression patterns after spontaneous differentiation were determined. For measuring effects on the first three endpoints, the cells were treated with various concentrations of MEHP dissolved in dimethyl sulfoxide (DMSO) and only with DMSO which served as control and harvested after 5 days. For measuring effects during spontaneous differentiation, the RNA of embryoid bodies (EBs) formed after 8 days' MEHP exposure was collected and changes in differentiation specific gene expression patterns were analyzed by quantitative real time RT-PCR.
RESULTSAs a result the viability and proliferation ability of both cell lines decreased significantly at 1000 µmol/L MEHP, while there was no effect on apoptosis or cell morphology. In addition MEHP also changed the gene expression pattern in the EBs of both cell lines.
CONCLUSIONMEHP in a high dose was cytotoxic and affected the development of hESCs, which indicates its embryo toxicity in human embryos.
Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Diethylhexyl Phthalate ; analogs & derivatives ; toxicity ; Dose-Response Relationship, Drug ; Embryonic Stem Cells ; drug effects ; pathology ; Humans
10.The species traceability of the ultrafine powder and the cell wall-broken powder of herbal medicine based on DNA barcoding.
Li XIANG ; Huan TANG ; Jin-le CHENG ; Yi-long CHEN ; Wen DENG ; Xia-sheng ZHENG ; Zhi-tian LAI ; Shi-lin CHEN
Acta Pharmaceutica Sinica 2015;50(12):1660-1667
Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall
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DNA Barcoding, Taxonomic
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drugs, Chinese Herbal
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analysis
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Phylogeny
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Plants, Medicinal
;
classification
;
genetics
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Powders
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Quality Control